OBJECTIVETo study the chemical constituents in the root of Isatis indigotica.

METHODThe constituents root were separated through various chromatographic techniques and their structures were elucidated by means of physicochemical properties and the analysis of their spectral data.

RESULTEleven compounds were isolated and identified as (+) -isolariciresinol (1), lariciresinol (2), lariciresinol-9-O-beta-D-glucopyranoside (3), lariciresinol-4'-O-beta-D-glucopyranoside (4), lariciresinol-4,4'-bis-O-beta-D-glucopyranoside (5), 3-formylindole (6), 1-methoxy-3-indolecarbaldehyde (7), 1-methoxy-3-indoleacetonitrile (8), deoxyvasicinone (9), epigoitrin (10), adenosine (11).

CONCLUSIONCompounds 4-8 were isolated from I. indigotica for the first time.

Furans ; analysis ; chemistry ; isolation & purification ; Glucosides ; analysis ; chemistry ; isolation & purification ; Indoles ; analysis ; chemistry ; isolation & purification ; Isatis ; chemistry ; Lignans ; analysis ; chemistry ; isolation & purification ; Lignin ; analysis ; chemistry ; isolation & purification ; Naphthols ; analysis ; chemistry ; isolation & purification ; Plant Extracts ; analysis ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo investigate the protective effect of lycopene against cryopreservation injury of post-thawing human sperm and its mechanism.

METHODSSemen samples were collected from 25 volunteers, each sample equally divided into four parts to be cryopreserved with cryoprotectant only (Ly0 control) or cryoprotectant + lycopene at the concentrations of 2 (Ly2), 5 (Ly5), and 10 µmol/L (Ly10), respectively. Before and after thawing, the semen samples were subjected to computer-assisted semen analysis ( CASA) for sperm kinematics, flow cytometry for sperm apoptosis, thiobarbituric acid assay for malondialdehyde (MDA) concentration, and JC-1 fluorescent staining for the sperm mitochondrial membrane potential (MMP).

RESULTSAfter cryopreservation, sperm motility was markedly decreased in all the groups (P < 0.01). The rate of sperm apoptosis was significantly lower in the Ly5 group than in the Ly0 control ([25.68 ± 4.36]% vs [33.26 ± 4.78]%, P < 0.05), while sperm MMP remarkably higher in the former than in the latter ([66.18 ± 14.23]% vs [55.24 ± 12.31]%, P < 0.05). The Ly2, Ly5 and Ly10 groups showed no statistically significance differences in the MDA level from the Ly0 control (P > 0.05).

CONCLUSIONAddition of lycopene at a proper concentration to cryoprotectant may reduce oxidative damage to sperm mitochondria in the freezing-thawing process, attenuate oxidative stress injury induced by reactive oxygen species to sperm plasma membrane, and improve the anti-apoptosis ability of sperm.

Apoptosis ; Carotenoids ; pharmacology ; Cryopreservation ; Cryoprotective Agents ; pharmacology ; Flow Cytometry ; Humans ; Male ; Malondialdehyde ; analysis ; Oxidative Stress ; Reactive Oxygen Species ; Semen Analysis ; Semen Preservation ; adverse effects ; methods ; Sperm Motility ; Spermatozoa ; drug effects ; physiology

To investigate the changes in bcl-2, bax expression and neuron apoptosis in the hippocampus after the blockade of cervical lymphatics, the model of lymphostatic encephalopathy was established by occluding and removing both the superficial and deep cervical lymph nodes in rats. The animals were sacrificed at 1, 2, 3, 5, 7 and 14 d after operation. H and E staining was used to observe the structure of brain tissues and TUNEL staining was used to detect in situ cell apoptosis in the hippocampus. The expression of bcl-2 and bax in the hippocampus were examined by RT-PCR. The results showed that cerebroedema appeared at day 2 and was most serious at day 5 after the blockade of cervical lymphatics. The number of TUNEL positive cells began to increase at day 2 and reached the maximum at day 5. The expression of bax began to increase at day 1 and reached the maximum at day 2. The expression of bcl-2 began to decrease at day 1 and dropped to the minimum at day 5. The items mentioned above recovered to control level at day 14. These results suggest that lymphostatic encephalopathy following the blockade of cervical lymphatics result in changes in bcl-2 and bax expression in the hippocampus and that apoptosis is the main form of neuron death.
Animals ; Apoptosis ; physiology ; Female ; Hippocampus ; metabolism ; pathology ; Lymph Node Excision ; Lymphatic System ; physiology ; Male ; Neck ; Neurons ; cytology ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; bcl-2-Associated X Protein ; genetics ; metabolism

OBJECTIVETo investigate the protective effect of astragalus on testis tissues following unilateral testicular torsion/detorsion.

METHODSThirty healthy adult Wistar rats were equally randomized into Group A (sham-operation control), B (torsion/detorsion) and C (torsion/detorsion plus intraperitoneal injection of astragalus). The testicular torsion/detorsion model was established by the Turner method. All the rats were fed under the same condition for 7 days and sacrificed, and the torsional testes were harvested for the detection of germ cell apoptosis, glutathione activity and the level of malonic diethylaldehyde (MDA).

RESULTSThe apoptosis indexes (AI) of spermatogenic cells in the torsional testes were (5.82 +/- 1.21), (36.18 +/- 8.40) and (20.39 +/- 3.57) in Group A, B and C, significantly higher in Group B and C than in A (P < 0.05) and in Group B than in C (P < 0.05). Significant differences were observed in glutathione activity in the ipsilateral testes among Group A (48.03 +/- 2.01), B (30.93 +/- 1.25), C (38.44 +/- 1.06) U/mg (P< 0.05), as well as in the level of MDA, (1.43 +/- 0.17), (3.98 +/- 0.36), (2.57 +/- 0.53) nmol/ml, among the three groups (P < 0.05).

CONCLUSIONAstragalus could significantly reduce the apoptosis of spermatogenic cells, decrease the level of lipid peroxidation and protect glutathione activity in the torsional testis.

Animals ; Apoptosis ; Astragalus Plant ; Disease Models, Animal ; Lipid Peroxidation ; Male ; Phytotherapy ; Rats ; Rats, Wistar ; Spermatic Cord Torsion ; drug therapy

BACKGROUNDTo study the relationship between hepatitis B virus genotyping Shenzhen isolates and HBV precore/core promoter mutation and antiviral effects.

METHODSThe HBV genotyping of 165 patients with HBV was carried out with mAbs ELISA. HBV precore/core promoter mutation was detected with gene chip technology in 24 patients with CHB. The relationship between HBV genotyping and interferon, lamivudine effects was analyzed.

RESULTS(1) Out of 165 cases, 106 (64.2%) of type B but 48 (29.1%) of type C were found. Type B accounted for 95.4% in group ASC, and type C for 64.7%in group LC (P<0.05). (2) Precore/core promoter mutation was found in 16 cases (10 of type B, and 6 of type C) out of 24 cases. Out of 16 cases, precore/core promoter mutation (nt1896, 1862) was found in 10 cases (9 cases of type B and 1 case of type C), while basal core promoter mutation (BCP mutation, nt1762,1764) was found in 6 cases (1 case of type B and 5 of type C). (3) Among 27 patients with CHB HBAg (+) treated with interferon, 11 cases of type B but 1 case of type C were tested to be fully responsive to interferon. Among 29 patients with CHB HBAg (+) treated with lamivudine, 15 cases of type B but 3 cases of type C were tested to be continuously responsive to lamivudine.

CONCLUSION(1) HBV genotype popularity in Shenzhen area was classified as type B the first and type C the second. (2) Type C seems more apt to develop BCP mutation and cirrhosis, and to be less responsive to interferon or lamivudine.

Adolescent ; Adult ; Aged ; Antiviral Agents ; therapeutic use ; Child ; Child, Preschool ; DNA, Viral ; genetics ; Female ; Genotype ; Hepatitis B ; drug therapy ; virology ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Interferons ; therapeutic use ; Lamivudine ; therapeutic use ; Liver Cirrhosis ; drug therapy ; virology ; Male ; Middle Aged ; Mutation ; Promoter Regions, Genetic ; genetics ; Treatment Outcome ; Viral Core Proteins ; genetics ; Young Adult

OBJECTIVEGastric cancer cells are insensitive to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). To sensitize gastric cancer cells to TRAIL, we treated gastric cancer MGC803 cells with TRAIL and cisplatin.

METHODSCell proliferation was measured using MTT assay. Cell apoptosis was determined by flow cytometry. Expression of proteins was analyzed by Western blot. The distribution of lipid rafts and death receptors was analyzed by immunofluorescence microscopy. MGC803 cells were pretreated with 50 mg/L nystatin for 1 h, and followed by the treatment of cisplatin and TRAIL.

RESULTS100 µg/L TRAIL resulted in (8.51 ± 3.45)% inhibition of cell proliferation and caused (3.26 ± 0.89)% cell apoptosis in MGC803 cells. Compared with the treatment with cisplatin alone, treatment with TRAIL (100 µg/L) and cisplatin (8.49 mg/L, IC(50) dose of 24 h) led to a dramatic increase in both inhibition of cell proliferation [(52.58 ± 4.57)% vs. (76.43 ± 5.35)%, P < 0.05] and cell apoptosis [(23.10 ± 3.41)% vs. (42.56 ± 4.11)%, P < 0.05]. Moreover, cleavage of caspase-8 and caspase-3 was detected. TRAIL (100 µg/L) did not induce obvious lipid rafts aggregation and death receptor 4 (DR4) clustering, while cisplatin (8.49 mg/L) significantly promoted the localization of DR4 in aggregated lipid rafts. Pretreatment with 50 mg/L nystatin, a cholesterol-sequestering agent, triggered (3.66 ± 0.52)% cell apoptosis after 24 h. Pretreatment with nystatin for 1 h before the addition of 8.49 mg/L cisplatin for 24 h caused a decreased tendency to cell apoptosis [(25.74 ± 3.28)% vs. (22.76 ± 2.97)%]. While, pretreatment with nystatin before the addition of cisplatin and TRAIL, the proportion of apoptotic cells decreased from (43.16 ± 4.26)% to (31.52 ± 3.99)% (P < 0.05).

CONCLUSIONCisplatin enhances TRAIL-induced apoptosis in gastric cancer MGC803 cells through clustering death receptors into lipid rafts.

Antineoplastic Agents ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cisplatin ; administration & dosage ; pharmacology ; Dose-Response Relationship, Drug ; Humans ; Membrane Microdomains ; metabolism ; Nystatin ; pharmacology ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology

Objective To investigate the expressions of human telomerase reverse transcriptase(hTERT) mRNA and protein in pheochromocytoma and paraganglioma and their significance as diagnostic markers in predicting the biological behaviour of these tumours.Methods Expression of hTERT mRNA was determined by in situ hybridization in 45 pheochromocytomas/paragangliomas(31 benign,7 suspected malignant and 7 malignant) and 9 normal adrenal medulla samples,hTERT protein was determined by immunohistoebemistry.Results hTERT mRNA was expressed in 5/7 malignant turnouts and 5/7 suspected malignant tumours as compared with 3/31 benign tumours(P

Objective To explore the problems in traditional Chinese medicine (TCM) scientific research project management and provide possible countermeasures.Methods Through cluster sampling,all the scientific research administrators and staff in a hospital were selected.A questionnaire about TCM hospital scientific research management was designed and used in our study.Results The majority of participants were basically satisfied with current scientific research management in TCM.Problems were analyzed including complex project application,singular evaluating standard,inflexible fund management,and backward professional concept.Conclusions The management of scientific research projects in TCM should combine professional characteristics,innovatively and comprehensively improve management concept,system,team and method.

Objective To investigate the effects of mercury on T lymphocytes and serum immune indexes of workers with Methods occupational mercury exposure. A total of 45 workers with occupational mercury exposure were selected as the ， mercury exposure group and 47 workers without occupational mercury exposure were selected as the control group using the judgment sampling method. Cold atomic absorption spectrometry was used to detect the urinary mercury level of the two groups. （ ） +， + +， + + - + Flow cytometry was used to detect the proportion of cluster of differentiation CD 3 CD3CD4 CD3CD8 and CD3CD19 ， - （ - ） - （ - ） cells in peripheral blood and the levels of tumor necrosis factor α TNF α and interleukin 8 IL 8 in serum. The levels of （ ） ， Results immunoglobulin Ig A IgG and IgM in serum were measured by immune nephelometry. The urinary mercury level of （ ： vs ，P ） individuals in the mercury exposed group was higher than that of the control group median 92.7 13.2 μg/g Cr <0.01 . The +， + +， - + proportion of CD3 CD3CD4 CD3CD19 cells in peripheral blood and serum IgG level in the mercury exposed group （ P ）， - - （ P ） decreased all <0.05 and the serum TNF α and IL 8 levels increased all <0.01 compared with the control group. Urinary - + mercury level was negatively correlated with the proportion of CD3CD19 cells in peripheral blood and serum IgG level in the ［ （r） ， ， P ］， study subjects Spearman correlation coefficient S were −0.21 and −0.31 respectively all <0.05 and positively - - （r ， ， P ） ， correlated with serum TNF α and IL 8 levels S were 0.36 and 0.39 respectively all <0.05 . However the urinary mercury （ P ）， +， + +， level was neither correlated with IgA and IgM levels in serum all >0.05 nor with the proportion of CD3 CD3CD4 + + （ P ） Conclusion CD3CD8 cells in peripheral blood all >0.05 . Occupational exposure to mercury can lead to abnormal ， changes in peripheral blood T lymphocyte subsets B lymphocytes and serum immune factors in workers. The mercury load of occupational mercury exposure workers may impact their immune function.

OBJECTIVETo study the dynamic changes of T lymphocyte subsets of AIDS patients during more than 24 months of highly active antiretrovirus therapy (HAART) with successful suppression of HIV replication and different CD4 + T cell restoration.

METHODSTotally 45 AIDS patients who had received HAART for more than 24 months were included. During HAART (including DO, M3, M6, M12, M18, and M24), the number of plasma HIV-1 RNA was measured quantitatively using the bDNA assay, and T lymphocyte subsets including CD3 + CD4 + cells, CD3 + CD8 + cells, naive CD4 + cells (CD4 + CD45RA + CD62L +), CD4 + CD28 + cells , and CD8 + CD38 + cells were detected with flow cytometer.

RESULTSAmong 45 patients, 24 patients (53.3%) whose plasma viral load decreased to less than 500 copies/ml at M6 and maintained to M24 were classified into three groups according to the CD4 + T cell count increments on M24 (compared with DO): group A (< 100/mm3), group B (100-200/mm3), and group C (> 200/mm3). After the initiation of HAART, T lymphocyte response, including CD4 + T cell counts, naive CD4 + cell counts, percentages of CD4 + CD28 + cells in these patients were improved gradually, while CD8 + CD38 + percentage decreased. The improvement of T lymphocyte response in group C was most remarkable even with highest plasma viral load and lowest CD4 T cell count on DO. Compared with group A and B, group C had significantly better improvement not only in the quantities of CD4 + T cell, but also in the CD28 + expression and naive CD4 + T cell populations.

CONCLUSIONST lymphocyte response of AIDS patients can be effectively reconstituted by HAART. Different dynamics of CD4 + CD28 + and naive CD4 + populations may considerably contribute to the quantity and cellular function restoration of CD4 + T lymphocyte.

Acquired Immunodeficiency Syndrome ; drug therapy ; immunology ; Anti-HIV Agents ; therapeutic use ; Antiretroviral Therapy, Highly Active ; CD4-Positive T-Lymphocytes ; immunology ; HIV ; drug effects ; physiology ; Humans ; T-Lymphocyte Subsets ; Virus Replication ; drug effects